摘要
[目的]通过构建能够删除选择标记基因的表达载体,消除由非目的基因带来的影响,更好地研究转入基因的单一功能。[方法]利用Cre/LoxP位点特异性重组系统,对DsRed2-1载体进行改造,分别引入LoxP序列及多克隆位点序列,并引入TK基因进行负筛选,最终获得无目的基因的表达载体。[结果]诱导的载体片段在分子水平上发生了重组,载体转染细胞在细胞水平上也产生了特异性红色荧光。[结论]Cre/loxP系统的标记基因诱导删除体系切实可行,有着广泛的应用前景。
[Objective]This study was to construct an expression vector capable of excising selectable marker gene,further eliminating the effect of marker gene on the functional study of target gene.[Method]By using Cre/LoxP site-specific recombination system,DsRed2-1 vector was modified by introducing LoxP sequence and multiple cloning site sequence,then the TK gene was ligated into this vector for negative selection.[Results]The fragments introduced were recombined at the molecular level under induced condition,and the specific red fluorescence was also observed in the recombinant vector transfected cells at the cellular level.[Conclusion] It is feasible to use this Cre/loxP system to excise marker genes,showing a broad application prospect.
出处
《安徽农业科学》
CAS
2013年第7期2852-2856,共5页
Journal of Anhui Agricultural Sciences
基金
新疆生产建设兵团博士基金(2011BB014)
新疆农垦科学院引导计划(YYD201110)
